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171 Cards in this Set
- Front
- Back
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inserted toad gene into bacteria
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Boyer and Cohen
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first person to study biotechnology
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Boyer
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wanted to splice cancer genes with E. coli genes
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Berg
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drank milk containing bacteria to prove it was safe
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Brenner
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made bacteria to eat oil spills, got them patented
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Chakrabarty
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founded Genentech with Boyer
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Swanson
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tried to isolate the insulin gene, but only found the one for rats
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Gilbert
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joined Genentech to make the insulin gene
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Goeddel
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added agrobacteria to parts of leaves
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Horsch
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70% of processed food contains____
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genetically modified ingredients
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manipulating organisms to make useful products
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biotechnology
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manipulation of genes
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genetic engineering
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rings of DNA in bacteria
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plasmids
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plasmid + foreign DNA -->
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recombinant DNA
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recombinant DNA + bacteria cell -->
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recombinant bacterium
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production of multiple copies of a single gene
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gene cloning
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cut DNA at specific locations
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restriction enzymes
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each restriction enzyme recognizes a specific ____
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restriction site / DNA sequence
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why don't restriction enzymes cut up the bacteria's own DNA?
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it's protected with methyl groups
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most restriction sites are ___
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symmetrical
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how long is an average restriction site?
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4-8 nucleotides
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single stranded end of fragment
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sticky end
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____ hooks the sticky ends together
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DNA ligase
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the original plasmid used for cloning is called the ____
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cloning vector
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40% of plants have received ____ gene
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herbicide resistance
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plants have been made to resist ___, ____, and ___.
they also have enlarged ___, improved ___, and ___better. |
herbicides, insects, ripening/spoilage.
economical parts, nutrition, look |
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golden rice was given the gene for ____
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beta carotene
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how does golden rice prevent blindness?
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makes beta carotene, which the body converts to retinol/vitamin A
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if plants were given the genes of Rhizobium, what would be the benefit?
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plants could do nitrogen fixation themselves, instead of needing bacteria to do it.
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solving a problem using an organism
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bioremediation
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microorganisms can be used to extract and transform ____, and also to degrade toxic ___.
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heavy metals
organic compounds |
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a good genus that could possibly clean up oil spills
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pseudomonas
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bacteria can handle ____better than we can
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radioactive materials
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drugs that stop a specific reaction step that causes disease
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target drugs
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drug for chronic myelogenous leukemia
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Gleevec
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an organism that contains genes from two species
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transgenic organism
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transplant organisms are made to be ____, so the ID genes match
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MHC compatible
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best cells to use for gene fixing
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bone marrow (actively mitotic)
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disease that Bubble Boy had
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severe combined immunodeficiency (SCID)
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what's it called when the cells reject the new DNA?
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a vector problem
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most common genetic disorder in caucasians
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cystic fibrosis
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what causes CF?
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problem with chloride ion channel
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process by which DNA is carried from one organism to another, using a virus.
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transduction
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carries genetic material
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vector
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old way of getting insulin?
human growth hormone? |
farm animals
pituitary gland of cadavers |
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dissolves blood clots and reduces <3attack risk.
given at sign of stroke. |
tPA - tissue plasminogen activator
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4 tests for a stroke
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smile
tongue arms speak |
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2 reasons for gene cloning
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make gene copies (library development)
harvest protein product |
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bacteria's best defense against phages
2nd best |
restriction enzymes
methylated A and C bases |
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putting the recominant DNA (plasmid+foreign) back into the bacterial cell
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transformation
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enzymes that chop in middle of DNA
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endonucleases
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enzymes that attack 3' or 5' end of DNA
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exonucleases
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specific site where DNA is cut
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restriction site
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2 characteristics of restriction fragment
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3-8 letters, palindromic
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single stranded end of fragment
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sticky end
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the plasmid that's going to get foreign DNA
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cloning vector
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plasmid that has foreign DNA
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recombinant plasmid
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a useful restriction enzyme would/wouldn't make sticky ends
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would!
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when foreign DNA is added to bacterial plasmids, you're building a ______
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bacterial (plasmid) library
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when foreign DNA is added to viruses, you're building a ______
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phage library
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2 characteristics of good cloning vector
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gene for antibiotic resistance
marker gene |
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what does a marker gene do?
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tells you if DNA has been added or not, by the color of the colonies.
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gene for lactose breakdown
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lacZ
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gene for ampicillin resistance
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amp^r
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Process of DNA being taken up by an organism
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Transformation
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How many people are carriers of CF?
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1 in 25
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What fraction of plasmids actually take up the DNA?
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1 in 10000
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Agar with something added
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Restrictive growth media
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How can you weed out the bacteria that have not taken up the ampicillin resistance gene?
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Grow them in agar with ampicillin, which will kill all non-resistant bacteria (that don't have the plasmid)
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Why would you see white colonies?
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The lacZ gene was cut and DNA inserted. Then lactose can't be used as food---> white
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Why would you see blue colonies?
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LacZ gene wasn't cut (no new DNA inserted) so galactose could still be used as a food source ----> blue
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Process of picking out ONLY the desired gene
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Nucleic acid hybridization
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Complementary sequence made in the lab to stick to the desired gene
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Probe
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How do you mark the probe sequence?
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Radioactive ACTG or fluorescent tag
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Where does the probe DNA go?
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It matches up with the complementary sequence found in the desired gene
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What do dark spots on the X-ray indicate?
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Colonies that definitely contain the desired gene (the probe stuck!)
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The library includes all colonies that have....
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Taken up the plasmid
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Before putting the foreign DNA into the plasmid, we have to...
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Get rid of unwanted DNA
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Enzyme that makes DNA from RNA
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Reverse transcriptase
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What does reverse transcriptase do?
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Makes cDNA complementary to the mRNA
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After the cDNA is made, what happens?
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Rnase is added for degradation- getting rid of all RNA
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Reverse transcriptase makes ____-stranded ___
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Single stranded DNA
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who invented DNA fingerprinting>
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Professor Sir Alec Jeffreys
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3 uses of DNA fingerprinting
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immigration arguments
criminal cases paternity tests |
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proof of inherited variation at DNA level
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restriction fragment length polymorphism
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a small DNA change that prevents enzymes from cutting
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single nucleotide polymorphism
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DNA with repeated sequence
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tandem repeat DNA
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piece of DNA that gets repeated
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minisatellite
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piece of DNA that is similar in many different minisatellites
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core sequence
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what replaced use of minisatellites in DNA profiling?
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polymerase chain reaction
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very short repeated sequences of DNA
(2 names) |
microsatellites/simple tandem repeats
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how many BP in a minisatellite?
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6-100
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how many BP in a microsatellite?
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1-7 bases
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how many times is a sequence repeated in a minisatellite?
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2-several hundred times
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how many times is a sequence repeated in a microsatellite?
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5-100 times
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where are minisatellites generally found in the double helix?
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toward the ends (telomeres)
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where are microsatellites generally found in the double helix?
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randomly scattered.
gotcha. |
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plasmid with powerful bacterial promoter sequence that the gene can be placed downstream of
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expression vector
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advantages of using yeast
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eukaryotic (can do RNA processing, has ER)
has plasmids too |
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plasmids made from yeast and bacterial DNA
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hybrid plasmids
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what's the advantage of artificial eukaryotic chromosomes?
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hundreds of genes can be added, instead of just a few into a plasmid (ring)
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3 requirements of artificial eukaryotic chromosomes
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centromere
telomeres origin of replication |
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changing genes that will affect future generations
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germ line therapy
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changing genes that will only affect the one organism
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somatic cell therapy
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artificial chromosomes could cause a ___ problem
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speciation
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2 techniques of making bacterial cells competent
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+2 ion salt water / heat shock
electroporation |
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example of a salt that could be used to increase competency
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CaCl_2
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opening up cell pores so that the cell takes up DNA more readily.
this process increases ____ |
competency
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3 ways of getting DNA into cells (not competency)
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use a bacteriophage (bacteria)
coat pellets w/ DNA and shoot into cells (plants) inject DNA w/ needle |
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hormone that regulates blood pressure
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angiotensin
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enzyme that makes a DNA copy of RNA
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reverse transcriptase
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enzyme used by HIV virus
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reverse transcriptase
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HIV is a ___
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retrovirus
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after reverse transcriptase builds the cDNA strand, _____makes the other side
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DNA polymerase
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study of what proteins are made by healthy/sick cells
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metabolomics
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method used to amplify DNA
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polymerase chain reaction
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reasons to use PCR
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DNA in bad shape
there isn't much DNA |
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inventor of PCR
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Kary Mullis
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what does taq mean/where does it come from?
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thermo aquaticus
from thermophile bacteria |
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unique enzyme for PCR
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taq polymerase
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state when repressor is bound to operator
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repressed
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state when repressor is not bound to operator
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induced
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how many cycles happen in PCR?
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30
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what do you put into the PCR machine?
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nucleotides, taq polymerase, targeted sequence, and primers
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how many molecules result from PCR?
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over 1 billion
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virus that was killing papayas
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ring spot virus
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gene that kills corn borer caterpillars
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BT gene
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what does the BT gene do?
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kill corn borer caterpillars
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why are GMOs risky?
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new proteins produced could cause allergic reactions
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type of corn eventually found in taco shells but was meant for animals
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StarLink
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species that has been affected by BT corn
why? |
monarch butterflies
they lay eggs on milkweed, which only grows near corn |
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a place where non-resistant pests can live (helps with resistance problem)
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refuge
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group that protested genetically modified technology
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Greenpeace
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group that started fires at MSU to protest GMO research
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Earth Liberation Front
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place that engineered salmon
(that grow __times faster) |
Aqua Bounty Farms
4 |
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2 types of gel for electrophoresis
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agarose and polyacrylamide
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positively charged electrode
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anode (attracts -)
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negatively charged electrode
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cathode (attracts +)
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2 things in buffer solution
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weak base (tris)
and its salt (boric acid) |
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it's best to use ___voltage to separate DNA
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low
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2 stains in electrophoresis
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methylene blue
ethidium bromide |
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electrophoresis separates molecules by ____
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size
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# bands in homo dominant
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1
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# bands in homo recessive
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2
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# bands in hetero
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3
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purpose of southern blot
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tell if DNA contains a certain sequence
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what is placed on gel in southern blot?
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nitrocellulose paper
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southern blot: ___solution pulled through gel into nitrocellulose paper
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basic
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after you have the nitrocellulose paper with DNA pattern on it, what do you do?
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add radioactive probe
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northern blots used for?
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RNA
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western blots used for?
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proteins
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what does RFLP stand for?
what are they? |
restriction fragment length polymorphisms
restriction fragments can have many different lengths |
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who came up with DNA fingerprinting?
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Sir Alec Jeffreys
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___regions of DNA are most polymorphic
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non-coding
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why are non-coding regions more polymorphic than coding regions?
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they are not conserved
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known regions that are highly polymorphic
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short tandem repeats
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faster method than Northern blotting
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reverse transcriptase-polymerase chain reaction (RT-PCR)
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"in place"
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in situ
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technique that uses labeled probes in the intact organism
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in situ hybridization
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what do you use to study a genome-wide expression
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DNA microarray assay
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technique of determining the function of a gene by disabling gene and observing consequences
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in vitro mutagenesis
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large scale analyses of genes of people with/without a condition
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genome-wide association studies
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a single base-pair site where variation is found in at least 1% of population
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single nucleotide polymorphism (SNP)
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what are used for ID markers in forensics?
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STRs
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who came up with the method for sequencing DNA?
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Sanger
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1st step of Sanger method
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take single stranded DNA and do PCR to amplify it
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to each sample of Sanger method, what do you add?
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primer complementary to 3' end
DNA polymerase all 4 bases as triphosphates |
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what does ddATP mean?
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di-deoxy adenine triphosphate
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what's unique about di-deoxy nucleotides?
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they're missing an oxygen (hydroxyl) off of the 3' carbon, as well as the 2' carbon
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in the Sanger method, what's the difference between what's added to each of the samples?
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#1 has ddATP
#2 has ddGTP #3 has ddCTP #4 has ddTTP |
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what gel is used in the Sanger method and why?
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polyacrylamide
it can separate strands that differ by only one base pair length |
